Introduction
Methods and materials
Data acquisition and preprocessing
Analysis of differentially expressed genes (DEG)
Enrichment analysis
Immune infiltration assay
Protein–protein interaction network (PPI)
Validation analysis
Construction of mouse subcutaneous tumor model
Cell cultures
Construction of lentiviral stably transfected cell lines
Western blot
RNA extraction and quantitative real-time polymerase chain reaction (qPCR)
Characteristic | Low expression of CXCL5 | High expression of CXCL5 | P |
---|---|---|---|
N | 84 | 84 | |
T stage, n (%) | 0.091 | ||
T1 | 46 | 44 | |
T2 | 23 | 25 | |
T3 | 10 | 9 | |
T4 | 5 | 6 | |
N stage, n (%) | 0.668 | ||
N0 | 79 | 80 | |
N1 | 5 | 4 | |
M stage, n (%) | 0.774 | ||
M0 | 81 | 79 | |
M1 | 3 | 5 | |
Pathologic stage, n (%) | 0.189 | ||
Stage I | 48 | 48 | |
Stage II | 21 | 23 | |
Stage III | 9 | 7 | |
Stage IV | 6 | 6 | |
Gender, n (%) | 0.598 | ||
Female | 42 | 40 | |
Male | 42 | 44 | |
OS event, n (%) | 0.014 | ||
Alive | 52 | 40 | |
Dead | 32 | 44 | |
Vascular invasion, n (%) | 0.077 | ||
No | 61 | 58 | |
Yes | 23 | 26 | |
Histologic grade, n (%) | 0.801 | ||
G1 | 23 | 20 | |
G2 | 48 | 49 | |
G3 | 11 | 12 | |
G4 | 2 | 3 | |
Age, median (IQR) | 65.8 (51, 66) | 66.1 (50, 73) | 0.401 |
AFP(ng/ml), median (IQR) | 8.3 (4, 108.5) | 25.86 (9,804.75) | < 0.01 |
BMI, median (IQR) | 26.24 (22.37, 28.47) | 22.56 (19.36, 31.01) | 0.029 |
Immunofluorescence and immunohistochemistry
Statistical analysis
Results
Elevated CXCL5 Expression in GBM
Patients exhibiting elevated CXCL5 expression experience an unfavorable prognosis
Identification of Differentially Expressed Genes (DEGs) Related to CXCL5 in GBM
Pathway enrichment analysis highlights CXCL5's Role in GBM
The correlation between CXCL5 expression and immune infiltration
The correlation between CXCL5 expression and clinical prognosis of GBM
Characteristics | Total (N) | Odds ratio (OR) | p-Value |
---|---|---|---|
T stage (T2 and T3 and T4 vs. T1) | 168 | 1.305(0.985–2.354) | 0.021 |
N stage (N1 vs. N0) | 172 | 3.447(1.568–3.889) | 0.386 |
N stage (N1 vs. N0) | 132 | 0.886(0.458–1.568) | 0.944 |
Pathologic stage (Stage III and Stage IV vs. Stage I and Stage II) | 128 | 1.251(0.986–2.033) | 0.158 |
Histologic grade (G3 and G4 vs. G1 and G2) | 176 | 1.748(1.048–2.355) | 0.018 |
Vascular invasion (Yes vs. No) | 158 | 1.528(1.088–2.417) | 0.048 |
AFP (ng/ml) (> 400 vs. ≤ 400) | 168 | 1.058(0.735–1.686) | 0.168 |
Albumin (g/dl) (≥ 3.5 vs. < 3.5) | 169 | 0.844(0.408–1.590) | 0.294 |
Tumor status (with tumor vs. tumor-free) | 172 | 1.568(1.120–2.322) | 0.318 |
Characteristics | Total (N) | Univariate analysis | Multivariate analysis | ||
---|---|---|---|---|---|
Hazard ratio (95% CI) | p-Value | Hazard ratio (95% CI) | p-Value | ||
Age | 284 | ||||
≤ 60 | 102 | Reference | |||
> 60 | 182 | 1.058(0.838–1.799) | 0.291 | 1.420 (0.845–2.565) | 0.334 |
Gender | 284 | ||||
Female | 134 | Reference | |||
Male | 150 | 0.755 (0.458–1.334) | 0.218 | 0.8585(0.418–1.708) | 0.899 |
Histologic grade | 286 | ||||
G1 | 41 | Reference | |||
G2 | 68 | 1.224 (0.586–1.996) | 0.598 | 0.825 (0.437–1.822) | 0.508 |
G3-G4 | 187 | 1.222 (0.710–2.103) | 0.484 | 1.041 (0.685–1.996) | 0.994 |
T stage | 248 | ||||
T1 and T2 | 158 | Reference | |||
T3 and T4 | 90 | 2.052 (1.661–3.241) | 0.014 | 2.255 (1.041–4.014) | 0.011 |
M stage | 268 | ||||
M0 | 264 | Reference | |||
M1 | 4 | 1.589 (1.007–3.288) | 0.012 | 1.085 (0.865–2.583) | 0.331 |
N stage | 258 | ||||
N0 | 253 | Reference | |||
N1 | 3 | 1.156 (0.855–2.609) | 0.344 | 1.186 (0.685–2.968) | 0.481 |
CXCL5 | 336 | 1.058 (0.755–1.969) | 0.004 | 1.035 (0.758–1.958) | 0.028 |
Pathologic stage | 308 | ||||
Stage I | 148 | Reference | |||
Stage II- IV | 160 | 1.256 (1.008–2.668) | 0.002 | 1.355 (0.868–2.055) | 0.314 |
Gene name | Forward | Reverse |
---|---|---|
CXCL5 | 5'-ATCGGCTACGTAGCTGATCG-3' | 5'-CGATCGACTAGCTTACGATG-3' |
CD68 | 5'-GCCATGACGTACGTGACTGC-3' | 5'-GCATGCTAGCATGCTAGCAT-3' |
MARCO | 5'-TACGTGCTAGCATGCATGCT-3' | 5'-AGCTACGTAGCTAGCTAGCT-3' |
IFN-γR1 | 5'-ATCGATCGATCGATCGATCG-3' | 5'-GATCGATCGATCGATCGATC-3' |